Raw data supporting: D-Enantiomeric β-Casein 197 Disrupts Biofilm-Associated Infection of MDR Pseudomonas aeruginosa and Acinetobacter baumannii through Superior LPS Binding and Host Cell Protection

This dataset contains all raw data supporting the findings reported in the manuscript: "D-Enantiomeric β-Casein 197 Disrupts Biofilm-Associated Infection of MDR Pseudomonas aeruginosa and Acinetobacter baumannii through Superior LPS Binding and Host Cell Protection."File descriptions:Biofilm_RawData.xlsx — Raw OD570 values from crystal violet biofilm inhibition and reduction assays against four P. aeruginosa (no. R1–no. R4) and four A. baumannii (no. 13–no. 16) clinical isolates. Seven of the eight isolates are MDR; P. aeruginosa no. R1 is β-lactam-resistant (n=5 per group).Cell viability_RawData.xlsx — Raw CCK-8 assay OD450 values for cytotoxicity evaluation of L- and D-β-casein 197 against A549 and J774A.1 cells at concentrations of 12.5, 25, 50, and 100 μM (n=3).FlowCyto_RawData.xlsx — Quadrant population percentages (Q1–Q4) from SYTO9/PI flow cytometry membrane integrity assays against all tested clinical isolates (n=3 per group).Flow_Cytometry_Raw_Plots.pdf — Representative raw dot plots from flow cytometry analysis showing gating strategy and quadrant populations.Galleria_RawData_Figshare.xlsx — Individual larval survival data from Galleria mellonella infection model (n=15 per group, 96 h observation, β-lactam-resistant P. aeruginosa no. R1 and MDR A. baumannii no. 13).ITC_RawData.xlsx — Raw isothermal titration calorimetry data (heat per injection, normalized heat, molar ratio) from three independent replicates of L- and D-β-casein 197 binding to LPS.LDH_CFU_RawData.xlsx — Raw OD490 values from LDH release assay and CFU counts from bacterial internalization assay in J774A.1 macrophages infected with MDR P. aeruginosa and A. baumannii (n=3 per group).LPS_neutralization_Raw_OD600_Data.xls — Raw OD600 readings from broth microdilution MIC assays in the LPS neutralization experiment (Fig. 7). Each compound (L-β-casein 197, D-β-casein 197, polymyxin B, gentamicin) was pre-incubated with exogenous P. aeruginosa LPS at 0, 1, 5, 10, 25, 50, and 100 μg/mL prior to MIC determination against P. aeruginosa no. R1 and A. baumannii no. 13. Three independent replicates per condition (n=3).Chymotrypsin_Stability_Raw_OD600_Data.xls — Raw OD600 readings from broth microdilution MIC assays in the proteolytic stability experiment (Supplementary Fig. S1). L- and D-β-casein 197 were incubated with chymotrypsin (1 mg/mL) at 37 °C and sampled at 0, 0.5, 1, 2, and 4 h, with residual antimicrobial activity assessed against P. aeruginosa no. R1 and A. baumannii no. 13. Three independent replicates per time point (n=3).TEM — Transmission electron microscopy original images of untreated, L-form-treated, and D-form-treated P. aeruginosa and A. baumannii clinical isolates.Biofilm CLSM.zip — Original confocal laser scanning microscopy images from biofilm inhibition and reduction assays stained with SYTO9/PI.Statistical_Analysis_RawData.xlsx — Supplementary statistical analysis file containing detailed information for all comparisons reported in the manuscript. For each figure, the worksheet lists the name of the statistical test used, degrees of freedom, and p-values reported to three decimal places (with p < 0.001 presented as <0.001). This file supports the statistical analyses described in the Methods section and the figure legends.All data are released under CC BY 4.0 licence.